LOCUS       sequence-430346-        6578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter cassette for SARGENT with piggyBac ITRs and 10x Capture 
            Sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6578)
  TITLE     Effect of genomic and cellular environments on gene expression noise
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6578)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pSARGENT1
FEATURES             Location/Qualifiers
     source          1..6578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(56..644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(136..155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(815..1606,1607..1675))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1438..1457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1676..1780)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2143..3325
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2374..3316
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3273..3293
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3333..3342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3339..3341,3342..3344,3345..3857)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(3384..3405)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3645..3664)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     repeat_region   3964..3998
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        4344..4647
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4648..4851
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             4876..5577
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     repeat_region   5881..5943
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     5976..5997
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6087..6208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6098..6117)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6152..6171
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
       61 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      121 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      181 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      241 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      301 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      361 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      421 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      481 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      541 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      601 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
      661 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      721 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      781 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      841 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      901 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      961 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1021 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1081 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1141 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1201 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     1261 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1321 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1381 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1441 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1501 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1561 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1621 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1681 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1741 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1801 caccttatac agttgaagtc ggaagtttac atacacttaa gttggagtca ttaaaactcg
     1861 tttttcaact actccacaaa tttcttgtta acaaacaata gttttggcaa gtcagttagg
     1921 acatctactt tgtgcatgac acaagtcatt tttccaacaa ttgtttacag acagattatt
     1981 tcacttataa ttcactgtat cacaattcca gtgggtcaga agtttacata cactaagttc
     2041 gactcctctg cagaatgcgg cgatgtttcg gtaaggggtc cgcgatcgcc ctaaggacgc
     2101 gtccgctctc gtcatcactg aggtggagaa gagcatgcgt gaggctccgg tgcccgtcag
     2161 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     2221 accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     2281 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     2341 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg
     2401 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac gcccctggct
     2461 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
     2521 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcttg ggcgctgggg
     2581 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
     2641 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
     2701 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
     2761 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
     2821 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
     2881 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
     2941 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
     3001 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
     3061 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
     3121 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
     3181 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
     3241 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
     3301 ttttcttcca tttcaggtgt cgtgaagtcg acgccaccat ggtgagcaag ggcgaggagc
     3361 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     3421 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     3481 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     3541 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     3601 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     3661 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3721 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     3781 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     3841 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     3901 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgcct
     3961 taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat attgctctct
     4021 ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg ccgcttggag
     4081 ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact ataacgaccg
     4141 cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta actcatacga
     4201 taattatatt gttatttcat gttctactta cgtgataact tattatatat atattttctt
     4261 gttatagata tcaggcgcgc caagcttgga tcctagtaat caattacggg gtcattagtt
     4321 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     4381 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     4441 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     4501 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     4561 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     4621 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     4681 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     4741 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     4801 acgcaaatgt gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     4861 aaccgtcaga ccggtatgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg
     4921 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac
     4981 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
     5041 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc
     5101 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca
     5161 tacgaggacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
     5221 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag
     5281 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa
     5341 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacatcaag
     5401 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg
     5461 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag
     5521 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct taattgagag
     5581 ctctcgcttc gagtctagag atcactcgag ttgtggccgg cccttgcttt aaggccggtc
     5641 ctagcaaagc tggtcggaag ttgggaaaac cctggcgtta aataaaatac gaaatgagcg
     5701 gccgctaaaa gttttgttac tttatagaag aaattttgag tttttgtttt tttttaataa
     5761 ataaataaac ataaataaat tgtttgttga atttattatt agtatgtaag tgtaaatata
     5821 ataaaactta atatctattc aaattaataa ataaacctcg atatacagac cgataaaaca
     5881 catgcgtcaa ttttacgcat gattatcttt aacgtacgtc acaatatgat tatctttcta
     5941 gggttaagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     6001 gccgccggga tcactctcgg catggacgag ctgtacaagt aataagttta aactcagtgt
     6061 ccatagcgac gccggcgcag acatgataag atacattgat gagtttggac aaaccacaac
     6121 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     6181 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     6241 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     6301 taattaagga ccccttaccg aaacatcgcc gcattctgca gaggagtcga gtgtatgtaa
     6361 acttctgacc cactgggaat gtgatgaaag aaataaaagc tgaaatgaat cattctctct
     6421 actattattc tgatatttca cattcttaaa ataaagtggt gatcctaact gacctaagac
     6481 agggaatttt tactaggatt aaatgtcagg aattgtgaaa aagtgagttt aaatgtattt
     6541 ggctaaggtg tatgtaaact tccgacttca actgtata
//
